Colored dissolved organic matter (CDOM) has been widely studied as part of efforts to improve understanding of the aquatic carbon cycle, by laboratory, in situ, and remote sensing methods. We studied ecoregion-scale differences in CDOM and dissolved organic carbon (DOC) to understand variability in organic matter composition and the use of CDOM as a proxy for DOC. Data from 299 lakes across the U.S. Upper Midwest showed that CDOM, measured as absorptivity at 440 nm (a440), correlated strongly with DOC (R2 = 0.81, n = 412). Colored lakes in the Northern Lakes and Forests (NLF) ecoregion drove this relationship. Lakes in the North Central Hardwood Forests (NCHF) had low color (most had a440 < 3 m-1) and weaker CDOM-DOC relationships (R2 = 0.47). Spectral slopes and specific ultraviolet absorbance (SUVA), indicated relatively low aromaticity and non-terrestrial DOM sources in low color lakes. Multiple regression analyses that included total dissolved nitrogen (TDN) and CDOM, but not chlorophyll a, improved DOC estimates in low color lakes, suggesting a dominant contribution of non-planktonic sources of low color DOM in these lakes. Our results show that CDOM is a reliable, regional proxy for DOC in lakes where forests and wetlands dominate the landscape and the DOM is primarily terrestrial in origin. Mapping of lake DOC at broad spatial scales by satellite-derived CDOM has lower accuracy in low color lakes.